While studying the nucleic acids of different strains of P. cinnabarinus (Pc58, Pc470.3, Pc470.6), the presence of dsRNA molecules (1.7 kb to 3.6 kb) was detected in two of them

CIFUENTES, V.

Martinez, C.

Pincheira, G.

English

Kansas State University

Fungal Genetics Reports Volume 37 Article 3 Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus V. CIFUENTES Universidad de Chile C. Martinez Universidad de Chile G. Pincheira Universidad de Chile Follow this and additional works at: https://newprairiepress.org/fgr This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. Recommended Citation CIFUENTES, V., C. Martinez, and G. Pincheira (1990) "Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus," Fungal Genetics Reports: Vol. 37, Article 3. https://doi.org/10.4148/1941-4765.1468 This Regular Paper is brought to you for free and open access by New Prairie Press. It has been accepted for inclusion in Fungal Genetics Reports by an authorized administrator of New Prairie Press. For more information, please contact cads@k-state.edu. Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus Abstract While studying the nucleic acids of different strains of P. cinnabarinus (Pc58, Pc470.3, Pc470.6), the presence of dsRNA molecules (1.7 kb to 3.6 kb) was detected in two of them. This regular paper is available in Fungal Genetics Reports: https://newprairiepress.org/fgr/vol37/iss1/3 Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus Cifuentes, V., C. Martinez and G. Pincheira Laboratorio de Genetica, Dpto. de Ciencias Ecologicas, Facultad de Ciencias, Universidad de Chile, Casilla 653, Santiago, Chile. While studying the nucleic acids of different strains of P. cinnabarinus (Pc58, Pc470.3, Pc470.6), the presence of dsRNA molecules (1.7 kb to 3.6 kb) was detected in two of them. Pycnoporus cinnabarinus is a basidiomycete capable of degrading lignocellulose and is known as a white rot fungus. P. cinnabarinus strains Pc58, Pc470.3 and Pc470.6 were isolated from carpophores collected in the rain forest of southern Chile. In the laboratory these strains were grown in liquid PC medium for 15 days at 30°C without shaking. The mycelia were harvested on Whatman #1 paper and washed with distilled water. Then the mycelia were frozen at -20°C and ground in a mortar. Nucleic acids were prepared by the rapid lithium chloride method (Leach et al. 1987 FGN 34:32-33), except that three phenol:chloroform:isoamyl alcohol (25:24:1) and two chloroform:isoamyl alcohol (24:1) extractions were performed before precipitating with ethanol. Nucleic acids were resuspended in sterile distilled water and later were dialyzed against TE buffer (10 mM Tris-HCl, 1 mM EDTA) for 16 h. The preparations were than analyzed by electrophoresis on a 0.7% agarose gel followed by staining with ethidium bromide. The electrophoretic analysis showed that strain Pc58 has only chromosomal DNA. Strains Pc470.3 and Pc470.6 have four bands moving faster than chromosomal DNA (Fig. 1). These bands correspond to dsRNA as determined by different tests with nucleases. These dsRNAs, denominated as L-dsRNA, M1-dsRNA, M2-dsRNA and S-dsRNA, have been purified by electrophoresis in low-melting-point agarose followed by CTAB extraction (Langridge et al. 1980 Anal. Biochem. 103:264-271). Their molecular sizes were approximately 3.6 kb, 2.7 kb, 2.3 kb and 1.7 kb respectively. P. cinnabarinus strains Pc470.3 and Pc470.6 seem to have the same types of dsRNA. Furthermore, the presence of different dsRNA in the same strain has been observed as in other fungi (Myers et al. 1988 Curr. Genet. 13:495-501). Since dsRNA is an indicator of the presence of viruses in fungi, our observations in P. cinnabarinus suggest the presence of a virus-like particle which does not seem to be associated with a visible alteration in phenotype of the fungus.  1CIFUENTES et al.: Presence of double stranded RNA in natural isolates of PycnoporusPublished by New Prairie Press, 2017 Figure 1. Agarose gel electrophoresis of nucleic acid preparations from natural isolates of P. cinnabarinus. Lanes 1 and 8: DNA digested with endonuclease HindIII; lane 2 strain Pc58; lane 4 strain Pc470.6 digested with DNAse I; lanes 5 and 6 strain Pc470.3; Lane 7 strain Pc470.6    2Fungal Genetics Reports, Vol. 37 [1990], Art. 3https://newprairiepress.org/fgr/vol37/iss1/3DOI: 10.4148/1941-4765.1468

Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus

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Presence of double stranded RNA in natural isolates of Pycnoporus cinnabarinus

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